The Focht Live-Cell Chamber System (FCS2)

Instructions

A message from the developer:
The FCS2 utilizes the most advanced technology for optimum functionality and versatility in the observation of living cells on a light microscope. The FCS2 was first manufactured in 1992. Since that time Bioptechs has improved and modified the system for a variety of specialized needs. As part of our dedication to respond to our customers needs we would be very appreciative of any comments and suggestions you may have relative to our products.

Controller User Guide

The controller operates on 120V-240V 50-60Hz. The AC power line input, power switch, voltage selector and fuses are all incorporated on the rear panel. The fuse carrier contains two 0.5A slow blow fuses. The fuses can be accessed by prying the fuse block out of its socket with a small screwdriver wedged under the small slot in the opening of the AC line receptacle. When replacing the fuse block make sure to align the arrow on the outside of the fuse block with the correct supply line voltage.
 

1. Turn the unit on using the main on-off switch located on the rear panel. All further adjustments are made on the front panel.
2. Set the display switch to the "SET" position. Insert the enclosed screwdriver into the hole marked Setpoint and adjust the controller to the desired setpoint.
3. Turn the display switch to the "Slide" or "Cmbr" position. The LCD will display a number between -1.8 and -4.2. This is normal.
4. Press the black button labeled "CALIBRATE" and make sure the display reads 26.6°C in the "Slide" position and 25.0 in the "Cmbr" position. If adjustment is necessary, turn the corresponding screws in the adjustment holes with the enclosed screwdriver. CW = increase, CCW = decrease.
5. Turn the controller off.
6. With the chamber loaded, plug the chamber into the controller. DO NOT plug the chamber into the controller unless it is either loaded or both slide contact wires are restricted from coming in contact with the base.

Special Notes

There is a safety cutout circuit that protects the cells if the microaqueduct drops below 1.0°C
from the setpoint after reaching setpoint minus 0.9°C. Only during a cold start will the safety cut-off circuit ignore the out-of-range condition. This safety circuit will shut off power to the slide circuit only and sound the alarm. If the alarm sounds, the user should correct the problem and press the red reset button located next to the red LED. This event can be the result of the following conditions:

1. Perfusion rate exceeds the rate which temperature stability can be maintained. To correct, press the red reset button as soon as the alarm sounds. This will reset the logic back to cold start up mode. Setpoint temperature will return within 15-20 seconds.
2. An interruption in AC line power may cause the logic to change state.
3. Physical displacement of the thermistor from the microaqueduct surface.
4. The thermistor that regulates the chamber frame is sealed internally eliminating accidental displacement.
   There is no alarm on the chamber frame circuit.

Chamber Loading

Your chamber has been inspected prior to packaging and is ready to use when you receive it. The chamber is shipped assembled and should be opened using the following procedure.

Rotate the large knurled ring in the base of the chamber clockwise until it stops. This will release and disengage the chamber top. Hold the white portion of the chamber while simultaneously (using thumb and forefinger) rotate the two smaller knurled nuts located on either side of the black electrical contact unit. This will remove the black portion which contains sensitive electronics so that they can remain dry. You now have the basic components of the chamber separated. Continue with the following directions;

Directions for Closed System use: (see drawing for part identification)
 

• Attach perfusion tubing to the inlet and outlet ports of the chamber. Bioptechs recommends 1/16" Tygon #2275 tubing for best biocompatability and ease of use. See perfusion drawing.
• Turn the top of the chamber (white) upside-down. Place the upper gasket (.75mm thick) into the recess while aligning the perfusion clearance holes with the perfusion tubes.
• Place the microaqueduct slide on the upper gasket gasket so that it is aligned with perfusion tubes.(grooved side up).
• Place the lower gasket (.1mm - 1mm thick) onto the microaqueduct slide and press around the perimeter creating a seal.
• Perfuse some media through the perfusion port producing a bead on the surface of the slide and displacing air trapped in the line.
• Lower the coverslip with cells onto the bead until it is resting on the gasket.
• Place the closure assembly on top of this stack while aligning the black electrical connector with the oval slot in the chamber top. Maintain a gentle pressure while turning the chamber over so that it is right side up. Then you can observe as the four paws engage the top through the depressions about the perimeter. Turn the large knurled ring counter-clockwise until tight. This will symmetrically tighten and seal the chamber.
• For applications involving rapid perfusion, a drop of immersion oil can be placed at the point where the base of the chamber meets the coverslip in the exposed aperture of the chamber. This drop will be drawn in by capillary action and enhance thermal conductivity to the coverslip. Place a light film of immersion oil on the flat surface of the surface probe to enhance thermal conductivity.
• Place the black power/sensor connector back on the two threaded posts and symmetrically rotate (using thumb and forefinger) the two knurled nuts onto the post. Make sure the spring wires make contact with the bus bars on the surface of the microaqueduct slide.
• It is important that the flat side of the surface probe is making uniform contact with the microaqueduct slide because the feedback loop that controls the microaqueduct temperature is reliant on this thermal contact point.
• Plug the FCS2 chamber into the controller 6 Pin mini DIN
• The display can be switched between Set, Slide and Cmbr at any time to monitor temperature without interfering with the regulation. If the chamber was preheated it will come up to temperature in about 1 minute. Otherwise it will take longer.
• The chamber should now be mounted onto the microscope adapter in the stage.
• The vertical portion of the drain tubing which extends down from the stage to your waste receptacle will create a siphon and form negative pressure in the chamber. This negative pressure will cause the coverslip to flex. This can be eliminated by breaking the siphon at a point equal to the height of the specimen with a "T" fitting.

Cleaning

The various components of the chamber and cleaning methods are as follows:

• The closure assembly and power/sensor connector do not come into contact with the specimen and should only be cleaned with a damp cloth or alcohol wipe. DO NOT immerse either one in liquid.
• Coverslips and gaskets should be hand cleaned and short-cycle autoclaved before use.
• The microaqueduct slide cannot be autoclaved. However, laboratory soap and/or alcohol an be used.
• The white plastic (Delrin) portion of the FCS2 will be in fluid communication with the specimen and can be autoclaved at 120°C for 15 min.